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1.
Several genes normally induced by estradiol (E(2)) in female fish, those for vitellogenins (VTGs) and zona radiata proteins (ZRPs), are also inducible in males exposed to estrogenic chemicals. Male sheepshead minnows (SHM) were exposed to both E(2) and para-nonylphenol (NP), at several doses and times to determine a dose-response. Quantitative real time PCR was used to measure mRNA for VTG1, VTG2, ZRP2 and ZRP3. Both E(2) and NP elicited a dose-response increase in all of the mRNAs tested. Exposure to both chemicals resulted in VTG2 expression at about a 10-fold lower level than VTG1, and ZRP2 expression at a lower level than ZRP3.  相似文献   

2.
Benzothiazole, a common chemical associated with tire manufacturing and industrial wastewater, is a principal component of both fresh water and estuarine tire leachate, a neurotoxicant to larval sheepshead minnows (Cyprinodon variegatus) in in vivo estuarine studies. The neurotoxic potential of benzothiazole was investigated following in vivo and in vitro exposure of sheepshead minnows to 3.75, 7.5, 15, 30 and 60 mg/l benzothiazole. Following benzothiazole exposure, fish were evaluated for survival, growth and histological alterations. Fish mortality occurred after 5 days of exposure to 60 mg/l (LC50 = 41.9). Significant decreases in larval growth were noted at all concentrations. Histologically, gills had cellular alterations but the central nervous system lacked the severe cellular damage seen in previous tire leachate exposure studies. Benzothiazole cytotoxicity to primary cultures of brain cells from sheepshead minnow and tilapia (Oreochromis niloticus) and two epithelial cell lines was evaluated using a tetrazolium salt assay (MTT) at 1 and 4 days. In vitro results indicate primary cultures of brain cells are less sensitive to benzothiazole than epithelial cell lines. Significant cytotoxicity to the epithelial cell lines was noted at 30 and 60 mg/l concentrations. Histologically and cytotoxicologically, the present study indicates that benzothiazole is a gill toxicant and not a neurotoxicant.  相似文献   

3.
Samples of used drilling muds collected during the course of a single well drilling operation exhibited different degrees of acute toxicity to sheepshead minnows and grass shrimp. For moulting grass shrimp, Palaemonetes pugio, the 96-h LC50's were 360 to 14 560 ppm (μl/litre); many of these values were considerably lower than those reported from previous drilling mud assays. However, when some of the muds used in this study were tested on sheepshead minnows, Cyprinodon variegatus, the resulting 96-h LC50's (6300 to 100 000ppm) were well within the range of previously reported values.Although a number of the drilling mud samples had relatively high amounts of chromium due to the addition of sodium chromate, there was a low correlation between chromium concentration and toxicity. In only three drilling muds could chromium content alone account for the observed toxicities. Furthermore, chemical analyses revealed the presence of No. 2 fuel oil like petroleum hydrocarbons in the mud samples. Based on the results of toxicity tests with No. 2 fuel oil and the concentrations of oil present in the muds, the toxicity of the mud samples to grass shrimp appears to be largely attributable to the petroleum hydrocarbon content.  相似文献   

4.
Several genes normally induced by estradiol (E2) in female fish, those for vitellogenins (VTGs) and zona radiata proteins (ZRPs), are also inducible in males exposed to estrogenic chemicals. Male sheepshead minnows (SHM) were exposed to both E2 and para-nonylphenol (NP), at several doses and times to determine a dose-response. Quantitative real time PCR was used to measure mRNA for VTG1, VTG2, ZRP2 and ZRP3. Both E2 and NP elicited a dose-response increase in all of the mRNAs tested. Exposure to both chemicals resulted in VTG2 expression at about a 10-fold lower level than VTG1, and ZRP2 expression at a lower level than ZRP3.  相似文献   

5.
Fathead minnows (Pimephales promelas) were exposed to the biocide chlorine dioxide (0.13 and 0.19 mg l-1) for up to 12 h and to its primary decomposition product, chlorite (177 and 304 mg l-1), for up to 96 h followed by recovery periods of up to 14 days. Chlorine dioxide exposure produced dose-dependent gill pathology including epithelial lifting, hypertrophy, hyperplasia, lamellar fusion, and necrosis. Complete recovery, even in fish with severe hypertrophy and lamellar fusion, was achieved within 4 days. Chlorite did not produce gill pathology even at a lethal exposure level (304 mg l-1 for 96 h) but did elicit a chronic inflammatory response with a marked increase in circulating and fixed phagocytes within hematopoietic and vascular tissues. This study indicates that chlorine dioxide is approximately 1000 times more toxic to fathead minnows than chlorite. Further, exposure of fathead minnows to these distinct but related compounds is consistently associated with very different pathologies.  相似文献   

6.
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7.
In vitro bioassays would facilitate monitoring of estrogen-like compounds in mussels (Mytilus galloprovincialis) since they tend to accumulate lipophilic compounds in their fat and muscle tissue. However, estradiol (E2) steroid (already identified in mussels) could induce false positive responses. This study focuses on temporal variability in estrogenic responses of mussel extracts and the possible relation of this response with E2 levels. The reproductive cycle and growth were thus monitored for six months in sexed mussels. E2 levels were similar between genders and these levels varied with reproductive stages. human estrogen receptor (hER) activities were similar at all stages except in February. E2 present in male and female extracts accounted for part of the hER activation observed, but no correlation was found between E2 levels and hER activities.  相似文献   

8.
Living barnacles eaten by sheepshead fishes results in the production of broken barnacle plate sediment. The yearly rate of production of broken barnacle plates is 4.9 kg m–2 and varies seasonally, with the largest mean flux in summer (22.5 g m–2 day–1) and the least in winter (1.9 g m–2 day–1). The mean grain size mode of broken barnacle plates is positively correlated with the flux of broken barnacle plates. Experiments with exclusion and inclusion cages support the postulation that increases in flux and size of broken barnacle plates are caused by the feeding activity of larger sheepshead.  相似文献   

9.
Ketoconazole is a fungicidal drug that inhibits function of cytochrome P450s in the synthesis of steroids. To examine if inhibition of P450 function affects gene expression in a dynamic manner, we conducted in vitro exposures of ovary tissue from fathead minnows (Pimephales promelas) to 0.5 microM ketoconazole to investigate effects on steroid production and gene expression over time. Expression of four key steroidogenesis genes was examined at 1, 6, and 12h of exposure. 11 beta- and 20 beta-hydroxysteroid dehydrogenases were down regulated at 1h and Cytochrome P450 17 was down-regulated at 12h, consistent with the absence of steroid production. In contrast, cytochrome P450 19A was up-regulated at 6h, indicating feedback regulation. Microarray analysis of 12h exposures indicated enrichment of biological processes involved in neurotransmitter secretion, lymphocyte cell activation, sodium ion transport, and embryonic development. These data suggest that, with the exception of cytochrome P450 19A, these steroid metabolic genes are regulated in a feed forward manner and that the effects of ketoconazole may be broader than anticipated based on the mechanism of action alone.  相似文献   

10.
To determine the estrogenicity of effluents from sewage treatment plants (STPs) to larval fish, 2-day-old sunshine bass were exposed to effluents from three STPs serving New York City (NYC), varying in size and treatment level. Estrogenic response was evaluated by measuring vitellogenin (VTG) and estrogen receptor (ER) expression in cytosolic fractions of whole body homogenates. Concentrations of the presumptive endocrine disruptors in the effluents were also measured. VTG and ER levels in sewage-exposed fish were 3-5 times that observed in controls. Combined concentrations of estradiol and estrone ranged from 5 to 13 ng/l and nonylphenol-ethoxylate metabolites (NPEOs: 4-nonylphenol, and 1-, 2-, and 3-nonylphenol-ethoxylates) ranged from 180 to 470 microg/l in chlorinated effluent. Results indicate that both ER and VTG can be used as biomarkers for endocrine disruption in larval fish, and that 4-day exposure to sewage effluent is sufficient to elicit significant expression of these markers in sunshine bass larvae. The extremely higher concentrations of NPEOs found in effluent relative to hormones (approximately 40,000-fold) indicates that surfactant metabolites may be contributing significantly to the estrogenic effects observed.  相似文献   

11.
Estrogens appear to have a modulating effect on the expression of cytochrome P4501A (CYP1A) in fish. A number of in vivo studies have demonstrated that hepatic CYP1A expression in females decrease during sexual maturation when plasma levels of 17 beta-estradiol (E2) increase, or in cases when the fish in injected with E2. Since a number of environmental contaminants have weak estrogen-like activities, the question arises if these compounds are able to modulate CYP1A expression as well. In the present study, we used in vitro monolayer cultures of rainbow trout, Oncorhynchus mykiss, liver cells to compare concentration-dependent (10(-9) to 10(-5) M) effects of the natural steroid E2 and the non-steroidal xenoestrogen 4-tert-octylphenol (OP) on CYP1A-catalyzed 7-ethoxyresorufin-O-deethylase (EROD) activity. The concentration dependency of the estrogenic activity of the two test compounds was assessed by determination of hepatocellular vitellogenin (Vg) release into the culture medium. Exposure of hepatocytes to E2 concentrations of 10(-8) M and higher led to a significant inhibition of basal cellular EROD activity. On the contrary, exposure to OP did not result in an inhibition of EROD activity, even at OP concentrations (10(-6) M, 10(-5) M) which were associated with a significant induction of Vg synthesis.  相似文献   

12.
The isomeric compositions of the eicosenoic and docosenoic fatty acids of four freshwater fish oils (from sheepshead Aplodinotus grunniens, tullibee Coregonus artedii, maria Lota lota and alewife Alosa pseudoharengus, respectively) were examined by open-tubular gas—liquid chromatography. The 22:1ω11 isomer, usually the dominant isomer in marine-fish oils, was unimportant relative to 22:1ω9. The unsaponifiables lacked fatty alcohols and pristane and were mostly cholesterol. The probability that the 22:1ω1 in marine fish oils is strictly of an exogenous origin (primarily originating as the fatty alcohol in copepod wax esters) is reviewed and it is proposed that docosenoic acids have no obligatory role in the lipid biochemistry of either marine or freshwater fish.  相似文献   

13.
Stress proteins (heat shock proteins, hsps) form part of the cellular protein repair system, and are induced by a wide variety of Stressors. To determine their suitability as tools for assessing sublethal sediment toxicity, we measured levels of members of the stress protein families hsp60 and hsp70 in benthic estuarine amphipods (Ampelisca abdita) exposed to sediments from 23 different sampling sites in San Francisco Bay for 10 d. Concentrations of sediment-associated xenobiotics were determined. Per cent survival was recorded and surviving animals were analysed for stress proteins using western blotting techniques. An inverse correlation (r2 = 0.44) was seen between amphipod survival and hsp64 levels, and hsp64 levels were positively correlated with concentrations of total polycyclic aromatic hydrocarbons (PAHs) (r2 = 0.5). Principal component analysis revealed that amphipod mortality was linked to a combination of several PAHs (phenanthrene, fluoranthene, pyrene, benzo(a)pyrene) and di-n-butylphthalate at southern San Francisco Bay sites. At northern San Francisco Bay sites, negative correlations were found between hsp64 levels and organotin compounds (MBT, DBT, TBT), and between hsp71 levels and the PAHs, benzo (b,k)fluoranthene and benzo(G,H,I)perylene, suggesting an inhibitory effect of these compounds on stress protein expression.  相似文献   

14.
Our goal was to study the involvement of cytochrome P450 genes in estrogen metabolism and the extent to which the potentially carcinogenic 4-hydroxyestradiol metabolite is formed by channel catfish (Ictalurus punctatus; CC). Estradiol metabolism and ethoxyresorufin-O-deethylase (EROD) activity were assessed in several tissues from fish collected from three variably contaminated sites in the Mississippi River Delta, from laboratory control fish, and from fish exposed to 20 mg/kg benzo(a)pyrene (BaP) i.p. for 4 days. Liver EROD activity was induced by BaP, but Delta fish EROD activities were not statistically higher than activities in control fish. Gill microsomal EROD activity was also induced by BaP, but activities were 8- to 77-fold lower than those from liver. The predominant estrogen metabolites formed by CC liver, gill and gonad microsomes were 2-hydroxyestradiol and estrone as detected by GC/MS. Liver and gill 2-hydroxyestradiol formation was induced in BaP-dosed fish. The trends in hydroxyestradiol formation in field collected fish were more variable. In all fish liver microsomes there was more 2- compared to 4-hydroxyestradiol formed. However, BaP-treatment increased the 4:2 hydroxyestradiol ratio from 0.04 in control fish to 0.2 in BaP-exposed fish, suggesting that BaP induces the formation of the potentially genotoxic estrogen metabolite. No detectable 4-hydroxyestradiol was produced by gill and gonad microsomes. These results will ultimately help in determining which fish P450 genes are susceptible to environmental contamination and may be involved in estrogen genotoxicity.  相似文献   

15.
Halogenated aromatic hydrocarbons (HAHs) and polynuclear aromatic hydrocarbons (PAHs) are major environmental contaminants. Fish species that are chronically exposed to these compounds can develop resistance to their toxic effects. In all fish species studied to date, toxicant resistance has been accompanied by decreased inducibility of the xenobiotic metabolizing enzyme, cytochrome P450 1A (CYPIA). CYP1A induction is mediated through the Aryl Hydrocarbon Receptor (AHR). Although these compounds mediate their effects through this pathway, there have been resistant populations in which one chemical class cannot induce CYPIA expression (HAHs) while the other (PAHs) can. Resistance to PAHs was examined in a HAH-resistant population of Fundulus heteroclitus collected from a site contaminated with both compound classes (Newark Bay, NJ). Fish were injected intraperitoneally with the HAH 3,4,3',4'-tetrachlorobiphenyl (PCB77), benzo[a]pyrene (B[a]P, a PAH) or vehicle and sacrificed after 2 (B[a]P) or 5 days (PCB77, vehicle). We found no significant increase in CYP1A mRNA levels in resistant Newark Bay F. heteroclitus treated with either B[a]P or PCB77, while there was a 3.9 fold (PCB77) and 4.2 fold (B[a]P) increase in CYP1A mRNA in Flax fish relative to controls. AHR labeling studies revealed significantly (P < 0.05) lower levels of hepatic AHR in Newark fish (1,770 +/- 1,693.2 DPM) relative to Flax fish (6,082.5 +/- 1,709.9 DPM). Overall, these data suggest Newark F. heteroclitus are resistant to both PAHs and HAHs at the level of CYP1A mRNA, which might be mediated, in part, though lower expression of AHR. We are currently studying the promoter sequence to determine its role in chemical resistance.  相似文献   

16.
As part of an effort to validate the use of heat shock proteins (hsp) as a biomarker of adverse effect, we investigated the impact of estrogen (E2), the hormone responsible for vitellogenisis in fish, on the induction of hsp 60 and 70. PLHC-1 (Pociliopsis lucida hepatoma cell line) cells were cultured under standard conditions and treated with either 16 ppm copper; a known inducer of hsp 60 and 70, 1 microM estradiol, a combination of copper and estradiol, or media alone. Copper induced both hsp 60 and 70 two fold, while E2 alone had no effect. The combination of the copper and E2 inhibited the hsp 70 response, indicated by a 51% decrease in cellular hsp 70 compared to the copper treatment alone. In contrast, the expression of hsp 60 in the cells exposed to the copper-estradiol mixture was not significantly different than that in the copper-exposed cells. The results suggest that the use of hsp 70 induction in fish liver as a biomarker of effect may be limited by the gender and sexual maturity of the fish.  相似文献   

17.
Environmental chemicals may modulate the endocrine system through interaction with plasma sex steroid-binding proteins (SBP) and SBP-regulated processes. Some of these chemicals, which are known to interact with the estrogen receptor (ER), were found to bind competitively to the Atlantic salmon (Salmo salar) SBP and potentially disrupt the endocrine function of these proteins. Furthermore, both weakly acting (di-n-butyl phthalate) and potent estrogen mimics (ethynylestradiol), were able to induce a substantial up-regulation of circulating levels of SBP in vivo. Interestingly, modulation of SBP-levels was found to be a more sensitive endpoint than chemically induced interference with classical ER-mediated mechanisms for weakly acting estrogen mimics like di-(n-butyl) phthalate. Interference with the endocrine function of SBPs may thus introduce a novel mechanism for endocrine disruption. and give additional answers to the question why some weakly acting xenoestrogens are causing "estrogen-like" reproductive disturbances in developing males.  相似文献   

18.
Environmental chemicals may modulate the endocrine system through interaction with plasma sex steroid-binding proteins (SBP) and SBP-regulated processes. Some of these chemicals, which are known to interact with the estrogen receptor (ER), were found to bind competitively to the Atlantic salmon (Salmo salar) SBP and potentially disrupt the endocrine function of these proteins. Furthermore, both weakly acting (di-n-butyl phthalate) and potent estrogen mimics (ethynylestradiol), were able to induce a substantial up-regulation of circulating levels of SBP in vivo. Interestingly, modulation of SBP-levels was found to be a more sensitive endpoint than chemically induced interference with classical ER-mediated mechanisms for weakly acting estrogen mimics like di-(n-butyl) phthalate. Interference with the endocrine function of SBPs may thus introduce a novel mechanism for endocrine disruption, and give additional answers to the question why some weakly acting xenoestrogens are causing “estrogen-like” reproductive disturbances in developing males.  相似文献   

19.
Several environmentally relevant contaminants are characterized as xenoestrogens by virtue of their ability to induce responses similar to 17β-estradiol (E2). There is concern that exposure to these xenoestrogens may result in endocrine and, thus, reproductive disruption. The objective of the present research was to determine whether xenobiotics known to elicit estrogenic responses in mammals were estrogenic in channel catfish, and if these compounds were capable of altering normal physiological response to E2. Sexually immature catfish were exposed by intraperitoneal injection to E2, suspected xenoestrogens or combination doses of E2 and xenoestrogen. Appearance of vitellogenin (Vg) in serum was used as the bioindicator of estrogenicity; Vg was measured by ELISA 7 days after injection. The ED50 of E2 for the appearance of Vg in blood, 0.6 mg/kg, was used as the positive control. Synthetic estrogens, ethinylestradiol, mestranol and DES were effective in inducing Vg; the antiestrogen tamoxifen inhibited response to E2 when given in a combination dose. Treatment by both methoxychlor and p-nonylphenol resulted in Vg appearance in serum; the doses required were 300 times that of E2, and the vitellogenic response was less when compared to the E2-induced response. In addition, a combination dose of methoxychlor with E2 decreased the magnitude of the response to E2. Other mammalian xenoestrogens, o,p′-DDT, chlordecone, lindane and β-HCH, were not able to produce a vitellogenic response in this study.  相似文献   

20.
Studies of xenobiotic metabolism and the regulation of enzyme systems for their metabolism (Phase I and II enzyme systems) require large numbers of animals, intensive use of experimental aquarium systems and in some instances can pose major problems when the compounds are scarce, expensive or too toxic to be disposed of easily. Previous studies have demonstrated the usefulness of isolated primary hepatocytes of fish for metabolic and enzyme induction studies. Functionally and structurally competent hepatocytes were isolated from juvenile plaice and after overnight acclimation to culture conditions, were exposed to a variety of PAHs for 24h. The levels of CYP1A1, phenol UDPGT and GST-A mRNAs were then estimated by slot blotting and hybridisation to their cDNA probes. The method enabled rapid and easy determination of the structure/activity relationships of these compounds as inducers of these key Phase I and II enzymes, and is potentially useful for screening large numbers of compounds both as an invitro toxicity test and for mechanistic studies.  相似文献   

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